LOCUS       sequence-365039-        6048 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low affinity version of the Serotonin Sensor sDarken under the 
            control of a hSyn Promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6048)
  AUTHORS   Kubitschke M, Muller M, Wallhorn L, Pulin M, Mittag M, Pollok S, 
            Ziebarth T, Bremshey S, Gerdey J, Claussen KC, Renken K, Gross J, 
            Gneisse P, Meyer N, Wiegert JS, Reiner A, Fuhrmann M, Masseck OA
  TITLE     Next generation genetically encoded fluorescent sensors for 
            serotonin.
  JOURNAL   Nat Commun. 2022 Dec 6;13(1):7525. doi: 10.1038/s41467-022-35200-w.
  PUBMED    36473867
REFERENCE   2  (bases 1 to 6048)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2022-12-6"; volume: "13(1):7525. doi"; pages: " 
            10.1038/s41467-022-35200-w"
COMMENT     Sequence Label: pAAV-SYN-L-sDarken
FEATURES             Location/Qualifiers
     source          1..6048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     142..161
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   320..449
                     /label=AAV2 ITR
     promoter        549..996
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      1009..1018
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1908..1929
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2035..2056)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2296..2315)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2719..3307
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2772..2792)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3190..3201)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3348..3555
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3725..3854
                     /label=AAV2 ITR
     rep_origin      complement(4091..4679)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4171..4190)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4850..5641,5642..5710))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5473..5492
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5711..5815)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(5841..6048,1..248))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5978..5999)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
       61 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
      121 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
      181 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
      241 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
      301 ctcttcgcta ttacgccagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
      361 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
      421 agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt
      481 atctacgtag ccatgctcta ggaagatcgt aggttaatta atctagactg cagagggccc
      541 tgcgtatgag tgcaagtggg ttttaggacc aggatgaggc ggggtggggg tgcctacctg
      601 acgaccgacc ccgacccact ggacaagcac ccaaccccca ttccccaaat tgcgcatccc
      661 ctatcagaga gggggagggg aaacaggatg cggcgaggcg cgtgcgcact gccagcttca
      721 gcaccgcgga cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg cctcagcact
      781 gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc cccttcccgg ccaccttggt
      841 cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga gatagggggg
      901 cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg
      961 ggcagcggag gagtcgtgtc gtgcctgaga gcgcagtcga gaggatccgc caccatggat
     1021 gtgctcagcc ctggtcaggg caacaacacc acatcaccac cggctccctt tgagaccggc
     1081 ggcaacacta ctggtatctc cgacgtgacc gtcagctacc aagtgatcac ctctctgctg
     1141 ctgggcacgc tcatcttctg cgcggtgctg ggcaatgcgt gcgtggtggc tgccatcgcc
     1201 ttggagcgct ccctgcagaa cgtggccaat tatcttattg gctctttggc ggtcaccgac
     1261 ctcatggtgt cggtgttggt gctgcccatg gccgcgctgt atcaggtgct caacaagtgg
     1321 acactgggcc aggtaacctg cgacctgttc atcgccctca acgtgctgtg ctgcacctca
     1381 tccatcttgc acctgtgcgc catcgcgctg gacaggtact gggccatcac ggaccccatc
     1441 gactacgtga acaagaggac gccccggcgc gccgctgcgc tcatctcgct cacttggctt
     1501 attggcttcc tcatctctat cccgcccatg ctgggctggc gcaccccgga agaccgctcg
     1561 gaccccgacg catgcaccat tagcaaggat catggctaca ctatctattc cacctttgga
     1621 gctttctaca tcccgctgct gctcatgctg gttctctatg ggcgcatatt ccgagctgcg
     1681 cgcttcctga gctcatataa gaacgtctat atcaaggccg acaagcagaa gaacggcatc
     1741 aaggcgaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgcctaccac
     1801 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1861 agcgtgcagt ccaaactttc gaaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1921 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gggcggtacc
     1981 ggagggagca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     2041 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg tgagggcgat
     2101 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     2161 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     2221 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacat ccaggagcgc
     2281 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     2341 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     2401 ctggggcaca agctggagta caactgtttt gaccaactgc gagagaggaa gacagtgaag
     2461 acgctgggca tcatcatggg caccttcatc ctctgctggc tgcccttctt catcgtggct
     2521 cttgttctgc ccttctgcga gagcagctgc cacatgccca ccctgttggg cgccataatc
     2581 aattggctgg gctactccaa ctctctgctt aaccccgtca tttacgcata cttcaacaag
     2641 gactttcaaa acgcgtttaa gaagatcatt aagtgtaagt tctgccgcca gtgagaattc
     2701 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     2761 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     2821 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     2881 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     2941 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3001 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3061 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     3121 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     3181 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     3241 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     3301 tccccgcatc gataccgtcg acccgggcgg ccgctcataa agctcgactg tgccttctag
     3361 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     3421 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     3481 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     3541 caggcatgct ggggatgcgg tccggactgt acaattgcat tcattttatg tttcaggttc
     3601 agggggagat gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg
     3661 ataaggatct tcctagagca tggctacgta gataagtagc atggcgggtt aatcattaac
     3721 tacaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     3781 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     3841 gagcgagcgc gcagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3901 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3961 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4021 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4081 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4141 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4201 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4261 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4321 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4381 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4441 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4501 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4561 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4621 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4681 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4741 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4801 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4861 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4921 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4981 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5041 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5101 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5161 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5221 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5281 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5341 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5401 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5461 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5521 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5581 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5641 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5701 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5761 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5821 ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     5881 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     5941 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     6001 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatca
//
